Macrophage-Specific NF-κB Activation Dynamics Can Segregate Inflammatory Bowel Disease Patients

• Published in: Frontiers in immunology 2019-09, Vol.10, p.2168-2168
• Main Authors: Papoutsopoulou, Stamatia, Burkitt, Michael D, Bergey, François, England, Hazel, Hough, Rachael, Schmidt, Lorraine, Spiller, David G, White, Michael H R, Paszek, Pawel, Jackson, Dean A, Martins Dos Santos, Vitor A P, Sellge, Gernot, Pritchard, D Mark, Campbell, Barry J, Müller, Werner, Probert, Chris S
• Format: Article
• Language: English
• Subjects: Active Transport, Cell Nucleus - genetics; Active Transport, Cell Nucleus - immunology; Adult; Animals; Cell Nucleus - genetics; Cell Nucleus - immunology; Colitis, Ulcerative - genetics; Colitis, Ulcerative - immunology; Colitis, Ulcerative - pathology; Crohn Disease - genetics; Crohn Disease - immunology; Crohn Disease - pathology; Crohn's disease; cytokines; Female; Humans; Immunology; inflammatory bowel disease; macrophages; Macrophages - immunology; Macrophages - pathology; Male; Mice; Mice, Knockout; Middle Aged; NF-κB; Signal Transduction - genetics; Signal Transduction - immunology; Transcription Factor RelA - genetics; Transcription Factor RelA - immunology; ulcerative colitis
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2019.02168

The heterogeneous nature of inflammatory bowel disease (IBD) presents challenges, particularly when choosing therapy. Activation of the NF-κB transcription factor is a highly regulated, dynamic event in IBD pathogenesis. Using a lentivirus approach, NF-κB-regulated luciferase was expressed in patient macrophages, isolated from frozen peripheral blood mononuclear cell samples. Following activation, samples could be segregated into three clusters based on the NF-κB-regulated luciferase response. The ulcerative colitis (UC) samples appeared only in the hypo-responsive Cluster 1, and in Cluster 2. Conversely, Crohn's disease (CD) patients appeared in all Clusters with their percentage being higher in the hyper-responsive Cluster 3. A positive correlation was seen between NF-κB-induced luciferase activity and the concentrations of cytokines released into medium from stimulated macrophages, but not with serum or biopsy cytokine levels. Confocal imaging of lentivirally-expressed p65 activation revealed that a higher proportion of macrophages from CD patients responded to endotoxin lipid A compared to controls. In contrast, cells from UC patients exhibited a shorter duration of NF-κB p65 subunit nuclear localization compared to healthy controls, and CD donors. Analysis of macrophage cytokine responses and patient metadata revealed a strong correlation between CD patients who smoked and hyper-activation of p65. These dynamic assays of NF-κB activation in blood-derived macrophages have the potential to segregate IBD patients into groups with different phenotypes and may therefore help determine response to therapy.